Kinetoplast DNA minicircles: regions of extensive sequence divergence.

Rogers, William O.; Wirth, Dyann F.

doi:10.1073/pnas.84.2.565

Cited by 52 publications

(27 citation statements)

References 23 publications

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“…16 This high sensitivity could be achieved because of the presence of kinetoplast minicircle DNA (thousands of copies per cell). 20 The specificity of LAMP is generally high because it uses four primers that recognize six locations on sample DNA.…”

Section: Discussionmentioning

confidence: 99%

Sensitive, Specific, and Rapid Detection of Leishmania donovani DNA by Loop-Mediated Isothermal Amplification

Takagi

Itoh

Islam³

et al. 2009

The American Journal of Tropical Medicine and Hygiene

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Abstract. We have applied a loop-mediated isothermal amplification (LAMP) technique to detect Leishmania donovani DNA. The LAMP technique detected 1 fg of L. donovani DNA, which was 10-fold more sensitive than a conventional polymerase chain reaction (PCR). All nested PCR-positive blood samples from visceral leishmaniasis patients were positive with the LAMP technique, and DNA samples from L. infantum , L. major , L. mexicana , L. tropica , L. braziliensis , Plasmodium falciparum , and healthy humans were negative with the LAMP technique. The advantages of the LAMP method are its shorter reaction time, a lack of requirement of sophisticated equipment, and visual judgment of positivity based on the turbidity of reaction mixture. Our LAMP technique can be a better alternative to a conventional PCR, especially under field conditions.

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Section: Discussionmentioning

confidence: 99%

Sensitive, Specific, and Rapid Detection of Leishmania donovani DNA by Loop-Mediated Isothermal Amplification

Takagi

Itoh

Islam³

et al. 2009

The American Journal of Tropical Medicine and Hygiene

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“…The most sensitive conventional diagnostic methods, culture of lesion biopsies and aspirates, are available only in reference laboratories. Even these less available, more sensitive methods are positive in only 70 to 75% of acute cases when optimally performed (21,38).Due to these difficulties several new approaches to the diagnosis of CL were developed, including DNA probes and PCR (15,29,31). Of these, the PCR method is the most promising (43).…”

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confidence: 99%

“…Of these, the PCR method is the most promising (43). The minicircle of kinetoplast DNA (kDNA) and ribosomal DNA are ideal targets for amplification because they are present in multiple copies and have both conserved and variable regions (7,18,31,36). Primers that amplify either segments of kDNA or the entire minicircle have great potential as diagnostic tools because they detect minute quantities of DNA and as little as 1/10 of a cultured Leishmania organism (7, 29).…”

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confidence: 99%

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PCR-Based Diagnosis of Acute and Chronic Cutaneous Leishmaniasis Caused by Leishmania ( Viannia )

et al. 2002

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We evaluated PCR methods for diagnosis of acute and chronic cutaneous leishmaniasis (CL) in an area of Colombia where Leishmania (Viannia) is endemic. The PCR method specifically amplified whole linearized minicircle kinetoplast DNA (kDNA) of the Leishmania subgenus Viannia from biopsy lysates. PCR products were detected in agarose gels. For 255 acute cases, this PCR method had greater sensitivity (75.7%) than each conventional method, i.e., microscopic examination of Giemsa-stained lesion scraping (46.7%), biopsy culture (55.3%), aspirate culture (46.3%), and the conventional methods combined (70.2%). Among 44 cases of chronic CL, amplification of biopsy DNA was more sensitive (45.5%) than the individual (4.5 to 27.7%) and combined (27.3%) conventional methods. The detection of kDNA in biopsies from chronic lesions was enhanced by a chemiluminescent dot blot hybridization, which produced a sensitivity of 65.8% when alone and 90.9% when in combination with DNA extraction of biopsy lysates (P < 0.001). Three biopsies from 84 skin lesions of other etiologies were falsely positive by PCR (specificity, 96.4%). PCR detected kDNA more frequently in biopsies (detection level, 83.9%) than in aspirates (74.7%) from 103 cases of acute CL. Among aspirates from 53 chronic cases of CL, the alternative methods, DNA extraction and hybridization, increased sensitivity from 41.5 to 56.6% (P > 0.05). This enhanced PCR method in chronic biopsies was so much more sensitive than conventional methods that it should be considered the preferred diagnostic method for chronic CL. These findings support the appropriate incorporation of PCR into diagnostic strategies for cutaneous leishmaniasis.The leishmaniases are a group of illnesses of the skin, oral and respiratory mucosae and the reticuloendothelium caused by protozoa of the genus Leishmania. Of these, the cutaneous form is the most widespread, afflicting primarily rural and periurban populations exposed to the infected sand fly vector. Cutaneous leishmaniasis (CL) is most frequently diagnosed by clinical evaluation, either alone or in combination with the leishmanin skin test. Clinical evaluation usually suffers from lack of standardization (13,38) and is hampered by the fact that even fairly typical acute lesions can be confused with other dermatological problems, such as sporotrichosis (9). The leishmanin skin test is highly sensitive but lacks specificity when employed in areas of endemicity because it cannot distinguish acute lesions from past infection. A definitive, laboratory diagnosis of mucosal or cutaneous leishmaniasis traditionally requires either the visualization of amastigotes or the isolation of replicative Leishmania from lesions (24). The most widely employed laboratory methods for CL are microscopic examination of lesion scrapings, biopsy impression smears, and histopathology. The most sensitive conventional diagnostic methods, culture of lesion biopsies and aspirates, are available only in reference laboratories. Even these less available, more sensitive methods ...

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“…The minicircle of kinetoplast DNA (kDNA) and ribosomal DNA are ideal targets for amplication because they are present in multiple copies and have both conserved and variable regions (de Bruijin and Barker, 1992;de Bruijin et al, 1993;Rogers and Wirth, 1994;Uliana et al, 1994). Several different methods are used for kDNA extraction.…”

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confidence: 99%

Cutaneous Leishmaniasis: Evaluation by Polymerase Chain Reaction in the Çukurova Region of Turkey

Ergïn

Erdoğan

Gümürdülü

et al. 2005

Journal of Parasitology

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The diagnosis of cutaneous leishmaniasis (CL) may depend on the detection of the parasite in histologic sections, the growth of the promastigotes in culture, or the identification of parasite by other techniques. We performed polymerase chain reaction (PCR) on paraffin-embedded biopsies to determine the validity of this technique for diagnosis of CL. PCR was used to detect the parasite using 2 different DNA extraction methods. PCR was positive in all 20 cases when the Leishmania parasite was detected by light microscopy. Twenty-seven of 34 cases that were negative microscopically for the parasite were positive using PCR. The first extraction method of DNA identified leishmanial DNA in 41 of 54 cases (75.9%); the second extraction of DNA was positive in 47 of 54 cases (87%). PCR was negative in all of the nonleishmaniasis cases. The PCR-based method appears to be a useful diagnostic approach for identification of suspected cases of CL.

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Kinetoplast DNA minicircles: regions of extensive sequence divergence.

Cited by 52 publications

References 23 publications

Sensitive, Specific, and Rapid Detection of Leishmania donovani DNA by Loop-Mediated Isothermal Amplification

Sensitive, Specific, and Rapid Detection of Leishmania donovani DNA by Loop-Mediated Isothermal Amplification

PCR-Based Diagnosis of Acute and Chronic Cutaneous Leishmaniasis Caused by Leishmania ( Viannia )

Cutaneous Leishmaniasis: Evaluation by Polymerase Chain Reaction in the Çukurova Region of Turkey

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